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Figure 5 | BMC Cancer

Figure 5

From: Glycogen synthase kinase-3 inhibition disrupts nuclear factor-kappaB activity in pancreatic cancer, but fails to sensitize to gemcitabine chemotherapy

Figure 5

Effects of gemcitabine combined with GSK-3 inhibition on NF-κB. A. Effect of AR-18 (50 μM, 8 h), gemcitabine (10 μM, 8 h), and their combination measured by NF-κB luciferase reporter assay. PANC-1 and MIA PaCa-2 cells were co-transfected with TA-LUC NF-κB reporter construct and β-gal (internal control) and then exposed to AR-18, gemcitabine or both. The normalized values are relative to the untreated control which is represented by dotted line (indicating basal level of NF-κB activity). Each column represents the mean for at least four experiments, each with three replicates; error bars = ± SEM. (*) significant: (p < 0.0003) when compared to untreated control. Gem: gemcitabine. B. Effect of genetic disruption of GSK-3 and its combination with gemcitabine on NF-κB activity measured by luciferase reporter assay. PANC-1 cells were genetically knocked down for GSK-3 isforms α(10 nM), β (80 nM) or both, and subsequently were co-transfected with TA-LUC NF-κB and β-gal (internal control) constructs. The genetically treated or untreated cells were then exposed to gemcitabine (10 μM, 8 h). The normalized values are relative to the untreated control which is represented by dotted line (indicating basal level of NF-κB activity). Each column represents the mean for at least four experiments, each with three replicates; error bars = ± SEM. (*) significant: (p = 0.08) when compared to untreated control. (**) significant: (p < 0.0005) when compared to untreated control. Gem: gemcitabine. Western blot analysis of expression of GSK-3α and β isoforms in the above cells confirms successful knock down of the target genes.

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